Article

Rapid neurotransmitter uncaging in spatially defined patterns. Nat Methods

Department of Molecular Biology, Lewis Thomas Laboratory, Washington Road, Princeton University, Princeton, New Jersey 08544, USA.
Nature Methods (Impact Factor: 25.95). 12/2005; 2(11):837-43. DOI: 10.1038/nmeth793
Source: PubMed

ABSTRACT Light-sensitive 'caged' molecules provide a means of rapidly and noninvasively manipulating biochemical signals with submicron spatial resolution. Here we describe a new optical system for rapid uncaging in arbitrary patterns to emulate complex neural activity. This system uses TeO(2) acousto-optical deflectors to steer an ultraviolet beam rapidly and can uncage at over 20,000 locations per second. The uncaging beam is projected into the focal plane of a two-photon microscope, allowing us to combine patterned uncaging with imaging and electrophysiology. By photolyzing caged neurotransmitter in brain slices we can generate precise, complex activity patterns for dendritic integration. The method can also be used to activate many presynaptic neurons at once. Patterned uncaging opens new vistas in the study of signal integration and plasticity in neuronal circuits and other biological systems.

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    • "The use of OAM for probing can lead to microscopic imaging with a spatial resolution that is higher than the Rayleigh limit Tamburini et al. (2006) and, when OAM fields are used in combination with suitable fluorescence methods (e.g., the stimulated emission depletion), they enable new methods of far-field microscopy with theoretically unlimited resolution Harke et al. (2006); Hell (2009). Optimal spatially structured light beams have also been considered as tools to cage/uncage specific molecules for accurate and rapid biological imaging Shoham et al. (2005). Some of these approaches may have relevant applications in the imaging of biological tissues, e.g. for diagnostic or research purposes. "
    Advanced Photonic Sciences, 03/2012; , ISBN: 978-953-51-0153-6
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    • "Over the last decade some new knowledge about neuronal-, genetic-, and quantumlevels of information processing in biological neural networks has been discovered [21], [28]. For example, whether a neuron spikes or not at any given time could depend not only on input signals and protein expression levels [9], [54], [77], but also on the physical properties of connections [28], on probabilities of spikes being received at the synapses [51], on neurotransmitters being secreted [30], [52], [76], on ion channels being opened, etc. [16], [24], [37], [45], [49]. As spiking processes in biological neurons are stochastic by nature, it would be appropriate to look for new ways to enhance the current SNN models with probabilistic parameters and to obtain probabilistic SNN models (pSNN). "
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    ABSTRACT: The paper proposes a novel research framework for building probabilistic computational neurogenetic models (pCNGM). The pCNGM is a multilevel modeling framework inspired by the multilevel information processes in the brain. The framework comprises a set of several dynamic models, namely low (molecular) level models, a more abstract dynamic model of a protein regulatory network (PRN) and a probabilistic spiking neural network model (pSNN), all linked together. Genes/proteins from the PRN control parameters of the pSNN and the spiking activity of the pSNN provides feedback to the PRN model. The overall spatio-temporal pattern of spiking activity of the pSNN is interpreted as the highest level state of the pCNGM. The paper demonstrates that this framework can be used for modeling both artificial cognitive systems and brain processes. In the former application, the pCNGM utilises parameters that correspond to sensory elements and neuromodulators. In the latter application a pCNGM uses data obtained from relevant genes/proteins to model their dynamic interaction that matches data related to brain development, higher-level brain function or disorder in different scenarios. An exemplar case study on Alzheimer's Disease is presented. Future applications of pCNGM are discussed.
    IEEE Transactions on Autonomous Mental Development 01/2012; 3(4-3):300 - 311. DOI:10.1109/TAMD.2011.2159839 · 1.35 Impact Factor
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    • "In these years, various methodological solutions have been adopted to optically stimulate neurons by caged compounds going from the use of UV sources (e.g. xenon flash lamps) coupled to the port of an epifluorescent microscope (Callaway & Katz, 1993), to the use of laser scanning approaches (Shoham et al., 2005) or digital holographic microscopes (Lutz et al., 2008). Moreover, also external devices such as optical fibres (Bernardinelli et al., 2005) or semiconductor UV light-emitting diodes (Venkataramani et al., 2007) have been used. "
    Optoelectronics - Devices and Applications, 10/2011; , ISBN: 978-953-307-576-1
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